Aryl cyclopropyl-amino-isoquinolinyl amide compounds

ABSTRACT

Provided herein are amino isoquinolinyl amide and sulfonamide compounds that affect the function of kinases and other proteins in a cell and that are useful as therapeutic agents. In particular, these compounds are useful in the treatment of eye diseases such as glaucoma and retinal diseases, as anti-inflammatory agents, for the treatment of cardiovascular diseases, and for diseases characterized by abnormal growth, such as cancers.

RELATED APPLICATIONS

This application claims the priority of U.S. Provisional PatentApplication No. 62/731,609, filed Sep. 14, 2018, and U.S. ProvisionalPatent Application No. 62/738,962, filed Sep. 28, 2018, the entirecontent of each of which is incorporated herein by reference.

SEQUENCE LISTING

This application contains a sequence listing having the filename1959002-00143_ST25.txt, which is 620 bytes in size, and was created onSep. 13, 2018. The entire content of this sequence listing is hereinincorporated by reference.

BACKGROUND

A variety of hormones, neurotransmitters and biologically activesubstances control, regulate or adjust the functions of living bodiesvia specific receptors located in cell membranes. Many of thesereceptors mediate the transmission of intracellular signals byactivating guanine nucleotide-binding proteins (G proteins) to which thereceptor is coupled. Such receptors are generically referred to asG-protein coupled receptors (GPCRs) and include, among others,α-adrenergic receptors, β-adrenergic receptors, opioid receptors,cannabinoid receptors and prostaglandin receptors. The biologicaleffects of activating or inhibiting these receptors is not direct, butis mediated by a host of intracellular proteins. The importance of thesesecondary proteins has been recognized and modulation of this class isnow being investigated as intervention points in disease states. One ofthe most important classes of these downstream effectors is the “kinase”class.

The various kinases play important roles in the regulation of variousphysiological functions. For example, kinases have been implicated in anumber of disease states, including, but not limited to: cardiacindications such as angina pectoris, essential hypertension, myocardialinfarction, supraventricular and ventricular arrhythmias, congestiveheart failure, atherosclerosis, renal failure, diabetes, respiratoryindications such as asthma, chronic bronchitis, bronchospasm, emphysema,airway obstruction, upper respiratory indications such as rhinitis,seasonal allergies, inflammatory disease, inflammation in response toinjury, rheumatoid arthritis. The importance of p38 MAPK inhibitors inparticular as new drugs for rheumatoid arthritis is reflected by thelarge number of compounds that has been developed over the last years(J. Westra and P. C. Limburg Mini-Reviews in Medicinal Chemistry Volume6, Number 8, August 2006). Other conditions include chronic inflammatorybowel disease, glaucoma, hypergastrinemia, gastrointestinal indicationssuch as acid/peptic disorder, erosive esophagitis, gastrointestinalhypersecretion, mastocytosis, gastrointestinal reflux, peptic ulcer,Zollinger-Ellison syndrome, pain, obesity, bulimia nervosa, depression,obsessive-compulsive disorder, organ malformations (e.g., cardiacmalformations), neurodegenerative diseases such as Parkinson's Diseaseand Alzheimer's Disease, multiple sclerosis, Epstein-Barr infection andcancer (Nature Reviews Drug Discovery 2002, 1: 493-502). In otherdisease states, the role of kinases is only now becoming clear. Theretina is a complex tissue composed of multiple interconnected celllayers, highly specialized for transforming light and color intoelectrical signals that are perceived by the brain. Damage or death ofthe primary light-sensing cells, the photoreceptors, results indevastating effects on vision. Despite the identification of numerousmutations that cause inherited retinal degenerations, the cellular andmolecular mechanisms leading from the primary mutations to photoreceptorapoptosis are not well understood, but may involve the wnt pathway (ASHackam “The Wnt Signaling Pathway in Retinal Degeneration” IUBMB LifeVolume 57, Number 6/June 2005).

The success of the tyrosine-kinase inhibitor STI571 (Gleevec) in thetreatment of chronic myelogenous leukemia (Nature Reviews Drug Discovery2003, 2: 296-313) has spurred considerable efforts to develop otherkinase inhibitors for the treatment of a wide range of other cancers(Nature Reviews Cancer 2003, 3: 650-665). The balance between theinitiation and the inactivation of intracellular signals determines theintensity and duration of the response of the receptors to stimuli suchas agonists. When desensitization occurs, the mediation or regulation ofthe physiological function mediated or regulated by the G proteins towhich the receptors are coupled is reduced or prevented. For example,when agonists are administered to treat a disease or condition byactivation of certain receptors, the receptors relatively quickly becomedesensitized from the action of the GRKs such that agonistadministration may no longer result in therapeutic activation of theappropriate receptors. At that point, administration of the agonist nolonger enables sufficient or effective control of or influence on thedisease or condition intended to be treated.

Janus Kinases (or JAK) are a family of cytoplasmic protein tyrosinekinases. The JAK family plays a role in the cytokine-dependentregulation of proliferation and function of cells involved in immuneresponse. Currently, there are four JAK family members are known JAK1,JAK2, JAK3, and TYK2. The JAKs usually associate with cytokine receptorsin pairs as homodimers or heterodimers. Specific cytokines areassociated with specific JAK pairings. Each of the four members of theJAK family is implicated in the signaling of at least one of thecytokines associated with inflammation. Binding of cytokine to aJAK-dependent cytokine receptor induces receptor dimerization whichresults in phosphorylation of tyrosine residues on the JAK kinase,effecting JAK activation. Phosphorylated JAKs, in turn, bind andphosphorylate various STAT proteins which dimerize, internalize in thecell nucleus and directly modulate gene transcription, leading, amongother effects, to the downstream effects associated with inflammatorydisease.

In view of the role that kinases play in many disease states, there isan urgent and continuing need for small molecule ligands which inhibitor modulate the activity of kinases. Without wishing to be bound bytheory, it is thought that modulation of the activity of kinases, inparticular ROCK and JAK kinases, by the compounds of the presentdisclosure is, at least in part, responsible for their beneficialeffects.

SUMMARY

In one aspect, provided herein are compounds of Formula (I):

or a pharmaceutically acceptable salt thereof,

wherein

R¹ is H, halo, —CN, —C₁₋₆ alkyl, —C₁₋₆ alkoxyl, —C₁₋₆ haloalkyl, —C₁₋₆haloalkoxyl;

R² is H or halo;

X is —(C₁₋₅ heteroaryl)—R³; and

R³ is halo, —C₂₋₆ heterocyclyl, or —C₁₋₆ heteroalkyl.

In one aspect, provided herein are compounds of Formula (Ia):

or a pharmaceutically acceptable salt thereof,

wherein

R¹ is H, halo, —CN, —C₁₋₆ alkyl, —C₁₋₆ alkoxyl, —C₁₋₆ haloalkyl, —C₁₋₆haloalkoxyl;

R² is H or halo;

X is —(C₁₋₅ heteroaryl)-(R³)_(n);

R³ is H, halo, —C₂₋₆ heterocyclyl, or —C₁₋₆ heteroalkyl; and

n is 1, 2, or 3.

In another aspect, provided herein are compounds of Formula (Ib):

or a pharmaceutically acceptable salt thereof,

wherein

R¹ is H, halo, —CN, —C₁₋₆ alkyl, —C₁₋₆ alkoxyl, —C₁₋₆ haloalkyl, —C₁₋₆haloalkoxyl;

R² is H or halo;

X is —(C₂₋₆ heterocyclyl)-(R³)_(n);

R³ is H, halo, —C₂₋₆ heterocyclyl, or —C₁₋₆ heteroalkyl; and

n is 1, 2, or 3.

In another aspect, provided herein are compounds having a structure:

or a pharmaceutically acceptable salt thereof.

In another aspect, provided herein are compositions, comprising acompound provided herein.

In another aspect, provided herein are pharmaceutical compositions,comprising a compound provided herein and a pharmaceutically acceptablecarrier.

In another aspect, provided herein are methods of treating an oculardisorder in a subject in need thereof, comprising administering to thesubject a therapeutically effective amount of a compound providedherein.

In another aspect, provided herein are methods of reducing intraocularpressure in a subject in need thereof, comprising administering to thesubject a therapeutically effective amount of a compound providedherein.

In another aspect, provided herein are methods of modulating kinaseactivity in a cell, comprising contacting the cell with a effectiveamount of a compound provided herein.

DETAILED DESCRIPTION

Publications and patents are referred to throughout this disclosure. AllU.S. Patent Applications, U.S. Patent Application Publications, and U.S.Patents referred to herein are hereby incorporated by reference in theirentirety. All percentages, ratios, and proportions used herein arepercent by weight unless otherwise specified.

Listed below are definitions of various terms used in the presentdisclosure. These definitions apply to the terms as they are usedthroughout this specification and claims, unless otherwise limited inspecific instances, either individually or as part of a larger group.

As used herein, the article “a” or “an” refers to one or to more thanone (i.e. to at least one) of the grammatical object of the article. Byway of example, “an element” means one element or more than one element.Furthermore, use of the term “including” as well as other forms, such as“include”, “includes,” and “included,” is not limiting.

As used herein, the term “administering” refers to administration of thecompounds provided herein to a cell or a subject as needed to achievethe desired effect.

As used herein, the term “alkoxyl” alone or in combination with otherterms means, unless otherwise stated, an alkyl group having thedesignated number of carbon atoms, as defined herein, connected to therest of the molecule via an oxygen atom.

As used herein, the term “alkyl” alone or in combination with otherterms means, unless otherwise stated, a straight or branched chainhydrocarbon having the number of carbon atoms designated (i.e., C₁₋₆means one to six carbon atoms) and includes straight or branched chainsubstituent groups.

As used herein, the term “composition” or “pharmaceutical composition”refers to a mixture of at least one compound—useful as describedherein—with a pharmaceutically acceptable carrier. The pharmaceuticalcomposition facilitates administration of the compound to a patient orsubject. Multiple techniques of administering a compound exist in theart including, but not limited to, intravenous, oral, aerosol,parenteral, ophthalmic, pulmonary, rectal, subcutaneous, and topicaladministration.

As used herein, the term “contacting a cell” is used to mean contactinga cell in vitro or in vivo i.e. in a subject, such as a mammal,including humans, rabbits, cats and dogs.

As used herein, the term “controlling the disease or disorder” is usedto mean changing the activity of one or more kinases to affect thedisease or disorder.

As used herein, the term “eye disease” includes, but is not limited to,glaucoma, allergy, cancers of the eye, neurodegenerative diseases of theeye, such as diabetic eye disease, macular degeneration (AMD),inflammation, and dry eye.

As used herein, the term “disease or disorder associated with kinaseactivity” refers to a disease, condition or disorder treatable, in wholeor in part, by inhibition of one or more kinases.

As used herein, the term “effective amount,” “pharmaceutically effectiveamount” or “therapeutically effective amount” refers to a nontoxic butsufficient dosage amount of an agent (e.g., the compounds orcompositions provided herein) to provide the desired biological result,which result may be reduction or alleviation, or both, of the signs,symptoms, or causes of a disease, or any other desired alteration of abiological system including influencing, reducing or inhibiting theactivity of or preventing activation of a kinase (e.g., modulatingkinase activity). An appropriate therapeutic amount in any individualcase may be determined by one of ordinary skill in the art using routineexperimentation. These terms as used herein may also refer to an amounteffective at bringing about a desired in vivo effect in an animal—wherein some embodiments, the animal is a human—including, but not limitedto, uveitis, reduction in intraocular pressure, or dry eye.

As used herein, the term “excipient” refers to physiologicallycompatible additives useful in preparation of a pharmaceuticalcomposition. Examples of pharmaceutically acceptable excipients (e.g.,pharmaceutically acceptable carriers) can, for example, be found inRemington Pharmaceutical Science, 16th Ed.

As used herein, the term “haloalkyl” refers to an alkyl groupindependently substituted with one or more (e.g., one to six) fluorine,chlorine, bromine, or iodine atoms. In some embodiments, the alkyl groupis independently substituted with one or more fluorine, chlorine, orbromine atoms. In some embodiments, the alkyl group is independentlysubstituted with one or more (e.g., one to three) fluorine or chlorineatoms.

As used herein, the term “halo” alone or in combination with other termsmeans, unless otherwise stated, halogen atoms such as fluorine,chlorine, bromine, or iodine atoms (e.g., F or CI).

As used herein, the term “haloalkoxyl” refers to an alkoxyl groupindependently substituted with one or more (e.g., one to six) fluorine,chlorine, bromine, or iodine atoms. In some embodiments, the alkoxylgroup is independently substituted with one or more fluorine, chlorine,or bromine atoms. In some embodiments, the alkoxyl group isindependently substituted with one or more (e.g., one to three) fluorineor chlorine atoms.

As used herein, the term “heteroalkyl” by itself or in combination withanother term means, unless otherwise stated, a stable straight orbranched chain alkyl group consisting of the stated number of carbonatoms and one, two or three heteroatoms independently selected from O,N, or S. The heteroatom(s) may be placed at any position of theheteroalkyl group, including between the rest of the heteroalkyl groupand the fragment to which it is attached, as well as attached to themost distal carbon atom in the heteroalkyl group.

As used herein, the term “heteroaryl” refers to a heterocyclic ringhaving aromatic character. In some embodiments, heteroaryl groups haveone to five carbon atoms. In some embodiments, heteroaryl groups havetwo to ten carbon atoms. In some embodiments, the heterocyclic ring is apolycyclic ring.

As used herein, the term “heterocyclyl” refers to a mono cyclicnon-aromatic radical, wherein the atoms forming the ring (i.e., skeletalatoms) include carbon atoms and one, two, three or four heteroatomsindependently selected from O, N, or S. In some embodiments, theheterocyclyl group is saturated or partially unsaturated.

As used herein, the term “subject,” “patient” or “individual” refers toa human or a non-human mammal. Non-human mammals include, for example,livestock and pets, such as ovine, bovine, porcine, canine, feline, andmurine mammals. In some embodiments, the patient, subject, or individualis human.

As used herein, the term “pharmaceutically acceptable” refers to amaterial that does not abrogate the biological activity or properties ofthe compound, and is relatively non-toxic, i.e. the material may beadministered to an individual without causing undesirable biologicaleffects or interacting in a deleterious manner with any of thecomponents of the composition in which it is contained.

As used herein, the term “pharmaceutically acceptable carrier” means apharmaceutically acceptable material, composition or carrier, such as aliquid or solid filler, stabilizer, dispersing agent, suspending agent,diluent, excipient, thickening agent, solvent or encapsulating material,involved in carrying or transporting a compound useful as providedherein within or to the patient such that it may perform its intendedfunction. Typically, such constructs are carried or transported from oneorgan, or portion of the body, to another organ, or portion of the body.Each carrier must be “acceptable” in the sense of being compatible withthe other ingredients of the formulation, including the compound usefulas provided herein, and not injurious to the patient. As used herein,“pharmaceutically acceptable carrier” also includes any and allcoatings, antibacterial and antifungal agents, and absorption delayingagents, and the like that are compatible with the activity of thecompound useful as provided herein, and are physiologically acceptableto the patient. The term “pharmaceutically acceptable carrier” mayfurther include a pharmaceutically acceptable salt of the compounduseful as provided herein. Other additional ingredients that may beincluded in the pharmaceutical compositions are described, for example,in Remington's Pharmaceutical Sciences (Genaro, Ed., Mack PublishingCo., 1985, Easton, Pa.), which is incorporated herein by reference. The“pharmaceutically acceptable carrier” is useful for the preparation of apharmaceutical composition that is: generally compatible with the otheringredients of the composition, not deleterious to the recipient, andneither biologically nor otherwise undesirable. “A pharmaceuticallyacceptable carrier” includes one or more than one carrier. Embodimentsinclude carriers for topical, ocular, parenteral, intravenous,intraperitoneal intramuscular, sublingual, nasal or oral administration.“Pharmaceutically acceptable carrier” also includes agents forpreparation of aqueous dispersions and sterile powders for injection ordispersions.

As used herein, the term “pharmaceutically acceptable salt” refers toderivatives of the compounds provided herein wherein the parent compoundis modified by converting an existing acid or base moiety to its saltform. Examples of pharmaceutically acceptable salts include, but are notlimited to, mineral or organic acid salts of basic residues such asamines; alkali or organic salts of acidic residues such as carboxylicacids; and the like. The pharmaceutically acceptable salts of thecompounds provided herein include the conventional non-toxic salts ofthe parent compound formed, for example, from non-toxic inorganic ororganic acids. The pharmaceutically acceptable salts of the compoundsprovided herein can be synthesized from the parent compound whichcontains a basic or acidic moiety by conventional chemical methods.Generally, such salts can be prepared by combining the free acid or baseforms of these compounds with a stoichiometric amount of the appropriatebase or acid in water or in an organic solvent, or in a mixture of thetwo; generally, nonaqueous media such as ether, ethyl acetate, ethanol,isopropanol, or acetonitrile may be used. Lists of suitable salts arefound in Remington's Pharmaceutical Sciences, 17th ed., Mack PublishingCompany, Easton, Pa., 1985, p. 1418 and Journal of PharmaceuticalScience, 66, 2 (1977), each of which is incorporated herein by referencein its entirety.

As used herein, the term “prevent” or “prevention” refers to no disorderor disease development if none had occurred, or no further disorder ordisease development if there had already been development of thedisorder or disease. Also considered is the ability of one to preventsome or all of the symptoms associated with the disorder or disease.

As used herein, the term “treatment” or “treating” refers to theapplication or administration of a therapeutic agent, i.e. a compoundprovided herein, to a patient, or application or administration of atherapeutic agent to an isolated tissue or cell line from a patient(e.g., for diagnosis or ex vivo applications), who has a disease, asymptom of the disease or the potential to develop the disease, with thepurpose to heal, alleviate, relieve, alter, remedy, ameliorate, improveor affect the disease, the symptoms of the disease, or the potential todevelop the disease. Such treatments may be specifically tailored ormodified, based on knowledge obtained from the field ofpharmacogenomics.

Compounds

In an aspect, provided herein are compounds of Formula (I):

or a pharmaceutically acceptable salt thereof,

wherein

R¹ is H, halo, —CN, —C₁₋₆ alkyl, —C₁₋₆ alkoxyl, —C₁₋₆ haloalkyl, or—C₁₋₆ haloalkoxyl;

R² is H or halo;

X is —(C₁₋₅ heteroaryl)—R³; and

R³ is halo, —C₂₋₆ heterocyclyl, or —C₁₋₆ heteroalkyl.

In another aspect, provided herein are compounds of Formula (Ia):

or a pharmaceutically acceptable salt thereof,

wherein

R¹ is H, halo, —CN, —C₁₋₆ alkyl, —C₁₋₆ alkoxyl, —C₁₋₆ haloalkyl, or—C₁₋₆ haloalkoxyl;

R² is H or halo;

X is —(C₁₋₅ heteroaryl)-(R³)_(n);

R³ is H, halo, —C₂₋₆ heterocyclyl, or —C₁₋₆ heteroalkyl; and

n is 1, 2, or 3.

In another aspect, provided herein are compounds of Formula (Ib):

or a pharmaceutically acceptable salt thereof,

wherein

R¹ is H, halo, —CN, —C₁₋₆ alkyl, —C₁₋₆ alkoxyl, —C₁₋₆ haloalkyl, or—C₁₋₆ haloalkoxyl;

R² is H or halo;

X is —(C₂₋₆ heterocyclyl)-(R³)_(n);

R³ is H, halo, —C₂₋₆ heterocyclyl, or —C₁₋₆ heteroalkyl; and

n is 1, 2, or 3.

In some embodiments, the compound of Formula (I) is a compound ofFormula (II):

or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula (I) is a compound ofFormula (III):

or a pharmaceutically acceptable salt thereof.

In some embodiments, R¹ is H, halo, —CN, —C₁₋₆ alkyl, —C₁₋₆ alkyl-OH,—C₁₋₆ alkoxyl, —C₁₋₆ haloalkyl, —C₁₋₆ haloalkoxyl;

In some embodiments, R¹ is H, halo, —CN, —C₁₋₃ alkyl, —C₁₋₃ alkoxyl,—C₁₋₃ haloalkyl, —C₁₋₃ haloalkoxyl.

In some embodiments, R¹ is H, Cl, —CN, —CH₃, —CH₂CH₃, —CH(CH₃)₂, —CHF₂,—CF₃, or —OCF₃.

In some embodiments, R¹ is Cl, —CN, —CH₃, —CH₂CH₃, —CH(CH₃)₂, —CHF₂,—CF₃, or —OCF₃.

In some embodiments, R¹ is H, Cl, —OH, —CH₂OH, —CN, —CH₃, —CH₂CH₃,—CH(CH₃)₂, —CHF₂, —CF₃, or —OCF₃.

In some embodiments, R¹ is Cl, —OH, —CH₂OH, —CN, —CH₃, —CH₂CH₃,—CH(CH₃)₂, —CHF₂, —CF₃, or —OCF₃.

In some embodiments, R² is F, Cl, Br, or I.

In some embodiments, R² is H or F.

In some embodiments, X is —(C₃₋₅ heteroaryl)-R³.

In some embodiments, X is —(C₃₋₅ heterocyclyl)-R³.

In some embodiments, X is a pyridyl, pyrrolyl, furanyl, thiophenyl,pyrazolyl, imidazolyl, pyrimidinyl, triazinyl, thiazolyl, pyrazinyl,pyridazinyl, or oxazolyl, each of which may be substituted with R³.

In some embodiments, X is

In some embodiments, X is

In some embodiments, X is

In some embodiments, X is -pyridinyl-(R³)_(n), -pyrimidinyl-(R³)_(n),-pyrazinyl-(R³)_(n), -pyridazinyl-(R³)_(n), or -thiazolyl-(R³)_(n).

In some embodiments, X is -tetrahydropyridinyl-(R³)_(n),-aziridinyl-(R³)_(n), -azetidinyl-(R³)_(n), -pyrrolidinyl-(R³)_(n),-piperidinyl-(R³)_(n), or -azepanyl-(R³)_(n).

In some embodiments, R³ is —C₂₋₆ heterocyclyl or —C₁₋₆ heteroalkyl.

In some embodiments, R³ is —C₃₋₅ heterocycloalkyl or —C₁₋₄ heteroalkyl.

In some embodiments, R³ is H or F.

In some embodiments, R³ is F, Cl, Br, or I.

In some embodiments, each R³ is, independently, F, Cl, Br, or I.

In some embodiments, n is 1. In some embodiments, n is 2.

In some embodiments, n is 2 and each R³ is, independently, F, Cl, Br, orI.

In some embodiments, R³ is F,

In some embodiments, X is

In some embodiments, X is

In some embodiments: R¹ is H, Cl, —CN, —CH₃, —CH₂CH₃, —CH(CH₃)₂, —CHF₂,—CF₃, or —OCF₃;

R² is H or F;

X is

and

R³ is F,

In some embodiments:

R¹ is Cl, —CN, —CH₃, —CH₂CH₃, —CH(CH₃)₂, —CHF₂, —CF₃, or —OCF₃;

R² is H or F;

X is

and

R³ is

In some embodiments, the compound is:

or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound is:

or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound is a compound of Table 1 or apharmaceutically acceptable salt thereof.

In some embodiments, the compound is a compound of Table 2 or apharmaceutically acceptable salt thereof.

Methods

In another aspect, provided herein are methods of treating an oculardisorder in a subject in need thereof, comprising administering to thesubject a therapeutically effective amount of a compound provided herein(i.e. a compound of Formula (I), Formula (Ia), Formula (Ib), Formula(II), Formula (III), Table 1, Table 2, or a pharmaceutically acceptablesalt thereof).

In some embodiments, the ocular disorder is glaucoma, an inflammatoryeye disease, a neurodegenerative eye disease, diabetic eye disease, wetage-related macular degeneration, or dry age-related maculardegeneration.

In another aspect, provided herein are methods of reducing intraocularpressure in a subject in need thereof, comprising administering to thesubject a therapeutically effective amount of a compound provided herein(i.e. a compound of Formula (I), Formula (Ia), Formula (Ib), Formula(II), Formula (III), Table 1, Table 2, or a pharmaceutically acceptablesalt thereof).

In another aspect, provided herein are methods of treating a disease ordisorder associated with a kinase activity in a subject in need thereof,comprising contacting the subject with a therapeutically effectiveamount of a compound provided herein (i.e. a compound of Formula (I),Formula (Ia), Formula (Ib), Formula (II), Formula (III), Table 1, Table2, or a pharmaceutically acceptable salt thereof).

In some embodiments, the kinase activity is a JAK (Janus kinase)activity.

In some embodiments, the kinase activity is a ROCK (Rho-associateprotein kinase) activity.

In some embodiments of these aspects, the compound is administeredtopically to an eye of the subject

In some embodiments of these aspects, the compound is administeredtopically to an eyelid of the subject.

In some embodiments of these aspects, the subject is a human.

In another aspect, provided herein are methods of modulating kinaseactivity in a cell, comprising contacting the cell with a effectiveamount of a compound provided herein (i.e. a compound of Formula (I),Formula (Ia), Formula (Ib), Formula (II), Formula (III), Table 1, Table2, or a pharmaceutically acceptable salt thereof).

In some embodiments, the cell is in a subject.

In some embodiments, the cell is in a human subject.

In another aspect, provided herein are uses of a compound providedherein (i.e. a compound of Formula (I), Formula (Ia), Formula (Ib),Formula (II), Formula (III), Table 1, Table 2, or a pharmaceuticallyacceptable salt thereof), a composition provided herein, or apharmaceutical composition provided herein, in the manufacture of amedicament for the treatment of a viral infection, a cancer, or anallergic disease.

In another aspect, provided herein are compositions, comprising acompound provided herein.

In another aspect, provided herein are pharmaceutical compositions,comprising a composition provided herein and a pharmaceuticallyacceptable carrier.

Actual dosage levels of an active ingredient in the pharmaceuticalcompositions provided herein may be varied so as to obtain an amount ofthe active ingredient that is effective to achieve a desired therapeuticresponse for a particular subject, composition, or mode ofadministration, without being toxic to the subject.

In some embodiments, it is especially advantageous to formulate thecompound in dosage unit form for ease of administration and uniformityof dosage. Dosage unit form as used herein refers to physically discreteunits suited as unitary dosages for the subject to be treated; each unitcontaining a predetermined quantity of therapeutic compound calculatedto produce the desired therapeutic effect in association with therequired pharmaceutical vehicle. The dosage unit forms of the presentdisclosure are dictated by and directly dependent on (a) the uniquecharacteristics of the therapeutic compound and the particulartherapeutic effect to be achieved, and (b) the limitations inherent inthe art of compounding/formulating such a therapeutic compound for thetreatment of the diseases referred to herein in a subject in needthereof.

In one embodiment, the compounds or compositions provided herein areformulated using one or more pharmaceutically acceptable excipients orcarriers. In one embodiment, the pharmaceutical compositions providedherein comprise a therapeutically effective amount of a compoundprovided herein and a pharmaceutically acceptable carrier.

In one embodiment, the present disclosure provides packagedpharmaceutical compositions comprising a container holding at least onetherapeutically effective amount of a compound provided herein, andinstructions for using the compound to treat one or more symptoms of adisease referred to herein in a subject in need thereof.

Routes of administration of any of the compositions provided hereininclude oral, nasal, rectal, intravaginal, parenteral, buccal,sublingual, topical, or ocular. The compounds for use as provided hereinmay be formulated for administration by any suitable route, such as forocular, oral or parenteral, for example, transdermal, transmucosal(e.g., sublingual, lingual, (trans)buccal, (trans)urethral, vaginal(e.g., trans- and perivaginally), (intra)nasal and (trans)rectal),intravesical, intrapulmonary, intraduodenal, intragastrical,intrathecal, subcutaneous, intramuscular, intradermal, intra-arterial,intravenous, intrabronchial, inhalation, and topical administration.

Suitable compositions and dosage forms include, for example, drops,tablets, capsules, caplets, pills, gel caps, troches, dispersions,suspensions, solutions, syrups, granules, beads, transdermal patches,gels, powders, pellets, magmas, lozenges, creams, pastes, plasters,lotions, discs, suppositories, liquid sprays for nasal or oraladministration, dry powder or aerosolized formulations for inhalation,compositions and formulations for ocular or intravesical administrationand the like. It should be understood that the formulations andcompositions that would be useful as provided herein are not limited tothe particular formulations and compositions that are described herein.

In another aspect, provided herein are dosage forms suitable foradministration to a subject in need thereof, comprising a compoundprovided herein (i.e. a compound of Formula (I), Formula (Ia), Formula(Ib), Formula (II), Formula (III), Table 1, Table 2, or apharmaceutically acceptable salt thereof).

In another aspect, provided herein are kits, comprising a compositionincluding a compound provided herein (i.e. a compound of Formula (I),Formula (Ia), Formula (Ib), Formula (II), Formula (III), Table 1, Table2, or a pharmaceutically acceptable salt thereof) and instructions foruse thereof. In some embodiments, the kit further includes one or moreof a syringe, a vial, or a dosage form.

Those skilled in the art will recognize, or be able to ascertain usingno more than routine experimentation, numerous equivalents to thespecific procedures, embodiments, claims, and examples described herein.Such equivalents were considered to be within the scope of thisdisclosure and covered by the claims appended hereto. For example, itshould be understood, that modifications in reaction conditions,including but not limited to reaction times, reaction size or volume,and experimental reagents, such as solvents, catalysts, pressures,atmospheric conditions, e.g., nitrogen atmosphere, and reducing oroxidizing agents, with art-recognized alternatives and using no morethan routine experimentation, are within the scope of the presentapplication.

It is to be understood that wherever values and ranges are providedherein, all values and ranges encompassed by these values and ranges,are meant to be encompassed within the scope of the present disclosure.Moreover, all values that fall within these ranges, as well as the upperor lower limits of a range of values, are also contemplated by thepresent application.

The following examples further illustrate aspects of the presentdisclosure. However, they are in no way a limitation of the teachings orpresent disclosure as set forth herein.

EXAMPLES

Compounds provided herein may be prepared as described in U.S. patentapplication Ser. No. 15/941,993.

Example 1: ROCK and JAK Assays

ROCK Kinase Inhibition Assays.

All compounds were initially prepared as 10 mM stocks in anhydrousdimethylsulfoxide (DMSO). A 20 μl aliquot of the 10 mM solutions wastransferred to individual wells in column 1 of a 96-well polypropylenemicrotiter plate (Corning #3363) and diluted with DMSO to give a finalcompound concentration of 4 mM. Test compounds were then seriallydiluted 1:5 in DMSO for an 11-point concentration response and furtherdiluted in the assay buffer bringing all compound concentrations to afinal range of 100 μM to 10 pM in 2.5% DMSO. The assay was performed inwhite 96-well, flat-bottom, half-area, non-binding assay plate (Corning#3642) in assay buffer consisting of 20 mM HEPES (pH 7.5), 10 mMMgCl₂*6H₂O, 100 μM sodium orthovanadate, 0.05% CHAPS and 0.1% bovineserum albumin. A 10 μL aliquot of compound from each well of theintermediate dilution plate and 20 μL of a 2× substrate/enzyme solutioncontaining acceptor substrate (800nM RSK2 peptide KRRRLSSLRA (SEQ ID NO:1)), ROCK2 enzyme (10 nM), or ROCK1 enzyme, and 1,4-Dithiothreitol (DTT,2 μM) were added to all wells. The reaction was initiated by theaddition of 10 μL of 4× stock solution ATP (2 ∥M). Reactions werethoroughly mixed manually, covered and allowed to incubate at roomtemperature for 75 min. Protein kinase activity was quantitated usingPromega's KINASE-GLO™ luminescent Kinase Assay Kit according to themanufacturer's directions. ATP concentrations remaining in Test wellsfollowing the termination of the enzymatic reaction were comparedagainst control wells containing equivalent amounts of DMSO containingno inhibitor (CTRL). ATP concentrations in both Test wells and CTRLwells were normalized against background (BKG) ATP concentrations inwells containing concentrations of inhibitor that completely inhibitedthe protein kinase under investigation (i.e. a concentration thatprevented any consumption of ATP over the course of the incubation).Percent of Control (POC) values were determined for each concentrationof compound tested according to the equation:

POC=((Test well value−BKG)/(CTRL−BKG))*100

IC₅₀ values were calculated using the following 4-parameter logisticcurve-fitting algorithm:

f(x)=(A+((B−A)/(1+((x/C){circumflex over ( )}D))))

IC₅₀ values were converted to K_(i) values using the Cheng-PrusoffEquation: K_(i)=IC₅₀/(1+([ATP]/Km ATP])).

JAK Kinase Assays.

Compounds were prepared in the exact same manner as described in theROCK Kinase Assay with the exception to the substrate and enzyme. TheJAK 2× substrate/enzyme solution consisted of acceptor substrate (800 nMAbl peptide EAIYAAPFAKKK (SEQ ID NO: 2)), JAK2 or JAK3 enzyme (10 nM)and DTT (2 μM). All other steps and solutions remain identical to theROCK Kinase Assay above. Results are shown below in Table 1 and Table 2.

Example 2: PTM-HTM Assay

Porcine Trabecular Meshwork cells (PTM) were isolated from freshlyobtained enucleated porcine eyes. Immortalized Human Trabecular Meshworkcells (TM-1) were obtained through a kind gift from Donna Peters in theDepartment of Ophthalmology and Visual Sciences at the University ofWisconsin. Cells were plated onto fibronectin coated glass-bottom96-well plates and allowed to attach overnight. Media was removed andreplaced with test compound in media with 1% fetal bovine serum andincubated for various times. After incubation, cells were formaldehydefixed, triton solubilized, and stained. PTM cells were stained withAlexa Fluor ®488 phalloidin (F-actin) and Hoechst 33342 (nuclei). TM-1cells were stained with anti-paxillin followed by Alexa Fluor ®488goat-anti-mouse IgG (focal adhesions) and Hoechst 33342 (nuclei). Allstaining reagents were obtained through Invitrogen. Images werecollected on an INCell 2200 imager with a 20× objective. The actin fiberlength and total area of focal adhesions were analyzed using customalgorithms developed in the INCell Developer Toolbox, v1.9.3. Datacollected were converted to percent of control (untreated cells). Curveswere fit to data in GraphPad Prizm using sigmoidal dose-response andconstraining top and bottom to 100% and 0%, respectively. Results areshown below in Table 1 and Table 2.

Example 3: Pharmaceutical Compositions for Lowering Intraocular Pressure

Topical pharmaceutical compositions of the compounds provided herein forlowering intraocular pressure are prepared by conventional methods. Acompound according to this disclosure is used as the free base or apharmaceutically acceptable salt thereof. When the composition istopically administered to the eyes once daily, the above compositiondecreases intraocular pressure in a subject suffering from glaucoma

Example 4: Pharmacological Activity for Glaucoma Assay

Pharmacological activity for glaucoma can also be demonstrated usingassays designed to test the ability of the subject compounds to decreaseintraocular pressure. Examples of such assays are described in thefollowing reference, incorporated herein by reference: C. Liljebris, G.Selen, B. Resul, J. Sternschantz, and U. Hacksell, “Derivatives of17-phenyl-18, 19, 20-trinorprostaglandin F_(2α) Isopropyl Ester:Potential Anti-glaucoma Agents”, Journal of Medicinal Chemistry 1995, 38(2): 289-304.

TABLE 1 Compound Structure IC₅₀ (nM)  1

  19 nM JAK2  148 nM JAK3   16 nM ROCK1   4.9 nM ROCK2  1130 nM IKK_(β) 2

 3

 4

 5

 6

  0.70 nM JAK2   1.5 nM JAK3  128 nM ROCK1   68 nM ROCK2   4.1 nMIKK_(β) PTM: 2100 nM   2.0 nM JAK1   0.25 nM TYK2  7

  1.0 nM JAK2   5.8 nM JAK3  1087 nM ROCK1  880 nM ROCK2   3.6 nMIKK_(β)  8

  0.65 nM JAK2   3.9 nM JAK3   7.3 nM IKK_(β)  121 nM ROCK1   93 nMROCK2  9

  1.0 nM JAK2   8.2 nM JAK3  1570 nM ROCK1  1165 nM ROCK2   14 nMIKK_(β) 10

  3.0 nM JAK2   14.0 nM JAK3  220 nM ROCK1   74 nM ROCK2  105 nMIKK_(β) >1000 nM Stat5   15 nM JAK1   34 nM TYK2 11

  10.8 nM JAK2  135 nM JAK3  153 nM ROCK1   15 nM ROCK2  765 nM IKK_(β)12

  17 nM JAK2  175 nM JAK3  759 nM IKK_(β)   21 nM ROCK1   21 nM ROCK2 13

  21 nM JAK2  179 nM JAK3  825 nM IKK_(β)   18 nM ROCK1   15 nM ROCK2 14

  0.90 nM JAK2   9.0 nM JAK3  122 nM ROCK1  108 nM ROCK2   16 nM IKK_(β) 200 nM Stat5  1080 nM PTM 15

  2.0 nM JAK2   4.8 nM JAK3  156 nM ROCK1   56 nM ROCK2   88 nM IKK_(β)16

16a

  2.0 nM JAK2   4.8 nM JAK3   56 nM ROCK2  156 nM ROCK1   88 nM IKK_(β)  0.8 nM JAK1   7.0 nM TYK2 16b

  1.0 nM JAK2   2.9 nM JAK3   25 nM ROCK2   74 nM ROCK1   70 nM IKK_(β)  0.44 nM JAK1   12 nM TYK2 17

  0.8 nM JAK2   4.2 nM JAK3   44 nM ROCK1   45 nM ROCK2 18

  2.5 nM JAK2   19.5 nM JAK3   6.4 nM ROCK1   4.0 nM ROCK2   17.5 nMIKK_(β) 19

 4975 nM JAK2  6750 nM JAK3  7400 nM ROCK1  4750 nM ROCK2    nM IKK_(β)20

  0.9 nM JAK2   2.4 nM JAK3   30 nM ROCK1   20 nM ROCK2   6.9 nM IKK_(β)21

22

23

24

25

26

27

  1.15 nM JAK2   19 nM JAK3  125 nM ROCK1   44 nM ROCK2  159 nM IKK_(β)  41 nM Stat5  120 nM JAK1   98 nM TYK2 28

 170 nM JAK1   1.4 nM JAK2   18 nM JAK3  160 nM ROCK1   67 nM ROCK2  381nM IKK_(β)   57 nM Stat5  210 nM TYK2 29

  43 nM ROCK1   13.8 nM ROCK2  170 nM JAK2  350 nM JAK3 >5000 nM IKK_(β)30

31

32

  0.35 nM JAK2   3.4 nM JAK3   5.0 nM IKK_(β)   20.0 nM ROCK1   12.3 nMROCK2 33

34

35

37

38

39

40

41

42

TABLE 2 Compound Structure IC₅₀ or Ki (nM) 36

  1.0 nM Ki ROCK2    6 nM Ki JAK2   211 nM Ki JAK3   1192 nM Ki IKK_(β)   12 nM IC₅₀ PTM 43

44

45

46

47

48

49

  2.0 nM JAK2    16 nM JAK3   3.2 nM IKK_(β)   181 nM ROCK2   207 nMROCK1    48 nM JAK1    91 nM TYK2 50

51

  3.0 nM JAK2    15 nM JAK3   132 nM IKK_(β)    8 nM ROCK2    9 nM ROCK1   11 nM JAK1    47 nM TYK2 52

  1.0 nM JAK2   4.4 nM JAK3   3.0 nM ROCK2   6.0 nM ROCK1    10 nMIKK_(β)   330 nM PKA   4.0 nM JAK1    15 nM TYK2 53

 0.95 nM JAK2   3.4 nM JAK3   3.2 nM IKK_(β)    35 nM ROCK2    67 nMROCK1   6.5 nM JAK1  12.7 nM TYK2 54

   12 nM JAK2   205 nM JAK3   3750 nM IKK_(β)    11 nM ROCK2    25 nMROCK1    17 nM JAK1   167 nM TYK2 55

  1.0 nM JAK2   4.8 nM JAK3    38 nM IKK_(β)   343 nM ROCK2   748 nMROCK1   0.9 nM JAK1   9.0 nM TYK2 56

  2.0 nM JAK2    12 nM JAK3   260 nM ROCK2   360 nM ROCK1    54 nMIKK_(β)    15 nM JAK1    16 nM TYK2 57

  370 nM ROCK2   700 nM ROCK1    14 nM JAK2   112 nM JAK3   1240 nMIKK_(β) 58

  385 nM ROCK2   915 nM ROCK1   225 nM JAK2   1430 nM JAK3   1630 nMIKK_(β) 59

  1.0 nM JAK2    13 nM JAK3   3.6 nM IKK_(β)   107 nM ROCK2   170 nMROCK1    28 nM JAK1    30 nM TYK2 60

 11.8 nM JAK2    13 nM JAK3   1585 nM IKK_(β)   5.9 nM ROCK2  15.5 nMROCK1    48 nM JAK1   110 nM TYK2 61

  108 nM JAK2   785 nM JAK3   5813 nM IKK_(β)   225 nM ROCK2   275 nMROCK1   465 nM JAK1   1065 nM TYK2 62

  2.0 nM JAK2   7.7 nM JAK3   6.4 nM IKK_(β)   116 nM ROCK2   190 nMROCK1    16 nM JAK1    15 nM TYK2 63

   72 nM JAK2   203 nM JAK3   320 nM IKK_(β)    37 nM ROCK2   116 nMROCK1   3300 nM JAK1   4200 nM TYK2 64

  250 nM JAK2   2476 nM JAK3   336 nM IKK_(β)    99 nM ROCK2   127 nMROCK1   1028 nM JAK1   2183 nM TYK2 65

66

   72 nM JAK2   1505 nM JAK3   319 nM IKK_(β)    39 nM ROCK2   116 nMROCK1   3344 nM JAK1   4278 nM TYK2 67

  2.0 nM JAK2  10.5 nM JAK3  11.5 nM IKK_(β)   1550 nM ROCK2   3400 nMROCK1  10.5 nM JAK1  19.5 nM TYK2 68

  3.0 nM JAK2    48 nM JAK3   9.1 nM IKK_(β)   3800 nM ROCK2   6500 nMROCK1   115 nM JAK1   145 nM TYK2 69

  3.0 nM JAK2    25 nM JAK3   370 nM ROCK2   470 nM ROCK1    16 nMIKK_(β)    53 nM JAK1    73 nM TYK2 70

  1.0 nM JAK2  14.5 nM JAK3   3.1 nM IKK_(β)   1830 nM ROCK2   3650 nMROCK1  24.4 nM JAK1    39 nM TYK2 71

 20.2 nM JAK2   200 nM JAK3   600 nM IKK_(β)   235 nM ROCK2   520 nMROCK1   120 nM JAK1   630 nM TYK2 72

  7000 nM JAK2  >50 k nM JAK3  >50 k nM ROCK2  >50 k nM RCK1 14,000 nMIKK_(β)  >50 k nM JAK1  >50 k nM TYK2 73

  5.9 nM JAK2    97 nM JAK3   1060 nM ROCK2   1216 nM RCK1  11.8 nMIKK_(β)   195 nM JAK1   230 nM TYK2 74

  7.4 nM JAK2    15 nM JAK3    25 nM ROCK2    18 nM ROCK1   250 nM JAK1  230 nM TYK2 75

76

   30 nM JAK2   725 nM JAK3    13 nM ROCK2    15 nM ROCK1   3600 nMIKK_(β)   140 nM JAK1   165 nM TYK2 77

   15 nM JAK2   380 nM JAK3   2350 nM IKK_(β)   8.0 nM ROCK2    14 nMROCK1    59 nM JAK1   250 nM TYK2 78

  2.0 nM JAK2    32 nM JAK3   4.0 nM IKK_(β)   220 nM ROCK2   310 nMROCK1    34 nM JAK1    55 nM TYK2 79

  7.0 nM JAK2    98 nM JAK3    31 nM ROCK2    68 nM ROCK1   208 nMIKK_(β)   110 nM JAK1   290 nM TYK2 80

  1.5 nM JAK2    15 nM JAK3    23 nM ROCK2    35 nM ROCK1   509 nMIKK_(β)   8.5 nM JAK1    40 nM TYK2 81

  4.4 nM JAK2    61 nM JAK3   500 nM IKK_(β)   3.0 nM ROCK2   5.1 nMROCK1    18 nM JAK1    51 nM TYK2 82

   13 nM JAK2   200 nM JAK3   507 nM IKK_(β)   4.5 nM ROCK2   9.0 nMROCK1   100 nM JAK1   189 nM TYK2 83

  9.4 nM JAK2    38 nM JAK3   280 nM IKK_(β)   3.5 nM ROCK2   6.4 nMROCK1    42 nM JAK1   112 nM TYK2 84

  3.4 nM JAK2    17 nM JAK3   235 nM IKK_(β)  17.3 nM ROCK2    46 nMROCK1 PTM: xx nM    16 nM JAK1    28 nM TYK2 85

  1.0 nM JAK2   5.8 nM JAK3   875 nM ROCK2   1200 nM ROCK1   3.6 nMIKK_(β)   7.3 nM JAK1    12 nM TYK2 86

  2.0 nM ROCK2   3.4 nM ROCK1   4.5 nM JAK2    23 nM JAK3  34.5 nMIKK_(β)    19 nM JAK1    79 nM TYK2 87

  0.8 nM JAK2   4.3 nM JAK3   2.7 nM IKK_(β)    45 nM ROCK2    44 nMROCK1    12 nM JAK1    19 nM TYK2 88

 10.8 nM JAK2   135 nM JAK3    57 nM ROCK2   153 nM ROCK1   765 nMIKK_(β)    68 nM JAK1   143 nM TYK2 89

   19 nM JAK2   148 nM JAK3   4.9 nM ROCK2    16 nM ROCK1   1130 nMIKK_(β)    95 nM JAK1   227 nM TYK2 90

  4975 nM JAK2   6750 nM JAK3   4750 nM ROCK2   7400 nM ROCK1 91

  1.0 nM JAK2   8.2 nM JAK3   1165 nM ROCK2   1570 nM ROCK1    14 nMIKK_(β)   5.0 nM JAK1   3.9 nM TYK2 92

  2.5 nM JAK2  19.5 nM JAK3   4.0 nM ROCK2   6.4 nM ROCK1  17.5 nMIKK_(β)    23 nM JAK1    74 nM TYK2 93

  0.9 nM JAK2   2.9 nM JAK3    20 nM ROCK2    30 nM ROCK1   6.9 nMIKK_(β)   2.0 nM JAK1   3.0 nM TYK2 94

 0.65 nM JAK2   3.9 nM JAK3   7.3 nM IKK_(β)    93 nM ROCK2   121 nMROCK1  0.80 nM JAK1    10 nM TYK2 95

   17 nM JAK2   175 nM JAK3   759 nM IKK_(β)    21 nM ROCK2    21 nMROCK1   102 nM JAK1   200 nM TYK2 96

 0.35 nM JAK2   2.4 nM JAK3   5.0 nM IKK_(β)  12.3 nM ROCK2  20.0 nMROCK1   3.0 nM JAK1   4.9 nM TYK2 97

98

   21 nM JAK2   179 nM JAK3   825 nM IKK_(β)    15 nM ROCK2    18 nMROCK1   121 nM JAK1   158 nM TYK2 99

 0.90 nM JAK2   9.0 nM JAK3   108 nM ROCK2   122 nM ROCK1    16 nMIKK_(β)  12.5 nM JAK1    11 nM TYK2 100

  1.0 nM JAK2   3.4 nM JAK3   147 nM ROCK2   250 nM ROCK1   6.0 nMIKK_(β)   1.0 nM JAK1   5.0 nM TYK2 101

  1.2 nM JAK2    18 nM JAK3    36 nM ROCK2   114 nM ROCK1   159 nMIKK_(β)    13 nM JAK1   146 nM TYK2 102

  3.0 nM JAK2  14.0 nM JAK3    74 nM ROCK2   220 nM ROCK1   105 nMIKK_(β)    15 nM JAK1    69 nM TYK2 103

 0.95 nM JAK2   1.9 nM JAK3   3.0 nM ROCK2   9.0 nM ROCK1   4.5 nMIKK_(β)   3.0 nM JAK1   5.0 nM TYK2 104

  1.5 nM JAK2    17 nM JAK3    67 nM ROCK2   160 nM ROCK1   381 nMIKK_(β)    8 nM JAK1    82 nM TYK2 105

  2.0 nM JAK2   1.9 nM JAK3   6.0 nM ROCK2   8.0 nM ROCK1   100 nMIKK_(β)    12 nM JAK1  0.70 nM TYK2 106

 0.70 nM JAK2   1.0 nM JAK3   3.0 nM ROCK2   4.7 nM ROCK1   3.2 nMIKK_(β)    12 nM PKC_(δ)  0.70 nM JAK1   1.5 nM TYK2 107

 0.70 nM JAK2   1.5 nM JAK3  0.95 nM JAK1   1.0 nM TYK2    68 nM ROCK2  128 nM ROCK1   4.1 nM IKK_(β) 108

 13.8 nM ROCK2    43 nM ROCK1   170 nM JAK2   350 nM JAK3  >5000 nMIKK_(β) 109

 12.4 nM ROCK2    28 nM ROCK1 JAK2 195 nM JAK3 695 nM 110

  1.5 nM JAK2   6.8 nM JAK3   7.9 nM ROCK2    13 nM ROCK1    15 nMIKK_(β)   456 nM PKA    27 nM JAK1   5.3 nM TYK2 111

  2.0 nM JAK2   4.8 nM JAK3    56 nM ROCK2   156 nM ROCK1    88 nMIKK_(β)   0.8 nM JAK1   7.0 nM TYK2 112

  1.5 nM JAK2   2.9 nM JAK3   194 nM ROCK2   470 nM ROCK1    12 nMIKK_(β)   0.8 nM JAK1   1.5 nM TYK2 113

  2.0 nM JAK2    24 nM JAK3    77 nM ROCK2   150 nM ROCK1    16 nMIKK_(β) 114

  2.0 nM JAK2   7.4 nM JAK3   265 nM ROCK2   533 nM ROCK1  10.0 nMIKK_(β)  11.0 nM JAK1   2.8 nM TYK2 115

  2.0 nM JAK2   6.3 nM JAK3    12 nM ROCK2    27 nM ROCK1   8.2 nMIKK_(β)    25 nM JAK1   7.0 nM TYK2 116

0.5-1.5 nM JAK2   1.9 nM JAK3   1.5 nM JAK1   2.9 nM TYK2   8.0 nM ROCK2   13 nM ROCK1    9 nM IKK_(β) 117

  4.9 nM ROCK2    10 nM ROCK1    21 nM JAK2   127 nM JAK3   144 nMIKK_(β) 118

  0.6 nM JAK2   4.7 nM JAK3   3.6 nM IKK_(β)   2.0 nM ROCK2   3.4 nMROCK1    25 nM JAK1    12 nM TYK2 119

   23 nM ROCK2    72 nM ROCK1    12 nM JAK2   118 nM JAK3   650 nMIKK_(β)    55 nM JAK1   1129 nM TYK2 120

   6 nM ROCK2    9 nM ROCK1   2.0 nM JAK2    10 nM JAK3    8 nM IKK_(β)121

   10 nM ROCK2    10 nM ROCK1  14.8 nM JAK2   153 nM JAK3   325 nM TYK2122

  7.0 nM ROCK2   8.2 nM ROCK1    59 nM JAK2   930 nM JAK3 JAK1 >500 nM123

  4.0 nM ROCK2   4.0 nM ROCK1 JAK2- 44 nM JAK3- 1140 nM 124

   31 nM ROCK2    48 nM ROCK1 JAK2 652 nM 125

   3 nM ROCK2   3.5 nM ROCK1 JAK2 65 nM 126

   5 nM ROCK2    5 nM ROCK1 JAK2 72 nM   900 nM JAK3 127

   9 nM ROCK2    25 nM ROCK1   900 nM JAK2   2000 nM JAK3   4100 nMIKK_(β) 128

   75 nM ROCK2   154 nM ROCK1    17 nM JAK2    36 nM JAK3 129

  1.0 nM JAK2   4.8 nM JAK3    38 nM IKK_(β)   343 nM ROCK2   748 nMROCK1   0.9 nM JAK1   9.0 nM TYK2 130

  1.0 nM JAK2   2.9 nM JAK3   120 nM ROCK2   250 nM ROCK1   5.0 nMIKK_(β)   1.0 nM JAK1  0.44 nM TYK2 131

  2.0 nM JAK2  25.0 nM JAK3    77 nM ROCK2   150 nM ROCK1    17 nMIKK_(β) 132

  1.5 nM JAK2   2.9 nM JAK3   194 nM ROCK2   470 nM ROCK1    12 nMIKK_(β) 133

 0.70 nM JAK2   1.5 nM JAK3    68 nM ROCK2   128 nM ROCK1   4.1 nMIKK_(β)   2.0 nM JAK1  0.25 nM TYK2 134

  2.0 nM JAK2   7.4 nM JAK3   265 nM ROCK2   606 nM ROCK1  10.0 nMIKK_(β)  11.0 nM JAK1   2.8 nM TYK2 135

   19 nM JAK2   148 nM JAK3   4.9 nM ROCK2    16 nM ROCK1   1130 nMIKK_(β) 136

  0.9 nM JAK2    15 nM JAK3    23 nM ROCK2    35 nM ROCK1   509 nMIKK_(β)   8.5 nM JAK1    40 nM TYK2 137

  7.0 nM JAK2    98 nM JAK3    31 nM ROCK2    68 nM ROCK1   208 nMIKK_(β) 138

139

140

 0.70 nM JAK2   1.5 nM JAK3    68 nM ROCK2   128 nM ROCK1   4.1 nMIKK_(β)   2.0 nM JAK1  0.25 nM TYK2 141

  1.0 nM JAK2   5.8 nM JAK3   875 nM ROCK2   1200 nM ROCK1   3.6 nMIKK_(β)   7.3 nM JAK1    12 nM TYK2 142

143

  3.0 nM JAK2  14.0 nM JAK3    74 nM ROCK2   220 nM ROCK1   105 nMIKK_(β)    15 nM JAK1    34 nM TYK2 144

 10.8 nM JAK2   135 nM JAK3    57 nM ROCK2   153 nM ROCK1   765 nMIKK_(β) 145

146

  2.0 nM ROCK2   3.4 nM ROCK1   4.5 nM JAK2    23 nM JAK3  34.5 nMIKK_(β) 147

   17 nM JAK2   175 nM JAK3   759 nM IKK_(β)    21 nM ROCK2    21 nMROCK1 148

149

150

151

  2.0 nM JAK2   7.4 nM JAK3   265 nM ROCK2   606 nM ROCK1  10.0 nMIKK_(β)  11.0 nM JAK1   2.8 nM TYK2 152

153

154

155

156

157

 0.90 nM JAK2   9.0 nM JAK3   108 nM ROCK2   122 nM ROCK1    16 nMIKK_(β) 158

  2.0 nM JAK2   6.3 nM JAK3  13.8 nM ROCK2    27 nM ROCK1   8.2 nMIKK_(β)    25 nM JAK1   7.0 nM TYK2 159

160

161

162

163

164

165

166

167

168

169

 1.15 nM JAK2    19 nM JAK3    44 nM ROCK2   125 nM ROCK1   159 nMIKK_(β)   120 nM JAK1    98 nM TYK2 170

 0.35 nM JAK2   3.4 nM JAK3   5.0 nM IKK_(β)  12.3 nM ROCK2  20.0 nMROCK1   3.0 nM JAK1   4.9 nM TYK2 171

172

173

174

  3.4 nM JAK2    17 nM JAK3   235 nM IKK_(β)  17.3 nM ROCK2   246 nMROCK1    16 nM JAK1    29 nM TYK2 175

   70 nM JAK2   200 nM JAK3   150 nM IKK_(β)   2.5 nM ROCK2   4.3 nMROCK1 PTM: 650 nM 176

  9.4 nM JAK2    38 nM JAK3   280 nM IKK_(β)   3.5 nM ROCK2   6.4 nMROCK1 177

178

179

180

While the disclosure has been described in detail and with reference tospecific embodiments thereof, it will be apparent to one skilled in theart that various changes and modifications can be made without departingfrom the spirit and scope of the disclosure.

1. A compound of Formula (I):

or a pharmaceutically acceptable salt thereof, wherein R¹ is H, halo,—CN, —C₁₋₆ alkyl, —C₁₋₆ alkoxyl, —C₁₋₆ haloalkyl, —C₁₋₆ haloalkoxyl; R²is H or halo; X is —(C₁₋₅ heteroaryl)-R³; and R³ is halo, —C₂₋₆heterocyclyl, or —C₁₋₆ heteroalkyl.
 2. The compound of claim 1, whereinthe compound of Formula (I) is a compound of Formula (II):

or a pharmaceutically acceptable salt thereof.
 3. The compound of claim1, wherein the compound of Formula (I) is a compound of Formula (III):

or a pharmaceutically acceptable salt thereof.
 4. The compound of claim1, wherein R¹ is H, halo, —CN, —C₁₋₃ alkyl, —C₁₋₃ alkoxyl, —C₁₋₃haloalkyl, —C₁₋₃ haloalkoxyl.
 5. The compound of claim 1, wherein R¹ isH, Cl, —CN, —CH₃, —CH₂CH₃, —CH(CH₃)₂, —CHF₂, —CF₃, or —OCF₃.
 6. Thecompound of claim 1, wherein R¹ is Cl, —CN, —CH₃, —CH₂CH₃, —CH(CH₃)₂,—CHF₂, —CF₃, or —OCF₃.
 7. The compound of claim 1, wherein R² is F, Cl,Br, or I.
 8. The compound of claim 1, wherein R² is H or F.
 9. Thecompound of claim 1, wherein X is —(C₃₋₅ heteroaryl)-R³;
 10. Thecompound of claim 1, wherein X is


11. The compound of claim 1, wherein R³ is —C₂₋₆ heterocyclyl or —C₁₋₆heteroalkyl.
 12. The compound of claim 1, wherein R³ is —C₃₋₅heterocycloalkyl or —C₁₋₄ heteroalkyl.
 13. The compound of claim 1,wherein R³ is F,


14. The compound of claim 1, wherein R¹ is H, Cl, —CN, —CH₃, —CH₂CH₃,—CH(CH₃)₂, —CHF₂, —CF₃, or —OCF₃; R² is H or F; X is

and R³ is F,


15. The compound of claim 1, wherein R¹ is Cl, —CN, —CH₃, —CH₂CH₃,—CH(CH₃)₂, —CHF₂, —CF₃, or —OCF₃; R² is H or F; X is

and R³ is


16. The compound of claim 1, selected from:

or a pharmaceutically acceptable salt thereof.
 17. A compound selectedfrom:

or a pharmaceutically acceptable salt thereof.
 18. A composition,comprising the compound of claim
 1. 19. A pharmaceutical composition,comprising the composition of claim 18 and a pharmaceutically acceptablecarrier.
 20. A method of treating an ocular disorder in a subject inneed thereof, comprising administering to the subject a therapeuticallyeffective amount of the compound of claim
 1. 21. The method of claim 20,wherein the ocular disorder is glaucoma, an inflammatory eye disease, aneurodegenerative eye disease, diabetic eye disease, wet age-relatedmacular degeneration, or dry age-related macular degeneration.
 22. Themethod of claim 20, wherein the compound is administered topically to aneye of the subject.
 23. The method of claim 20, wherein the compound isadministered topically to an eyelid of the subject.
 24. A method ofreducing intraocular pressure in a subject in need thereof, comprisingadministering to the subject a therapeutically effective amount of thecompound of claim
 1. 25. The method of claim 24, wherein the compound isadministered topically to an eye of the subject
 26. The method of claim24, wherein the compound is administered topically to an eyelid of thesubject.
 27. A method of modulating kinase activity in a cell,comprising contacting the cell with an effective amount of the compoundof claim
 1. 28. The method of claim 27, wherein the cell is in asubject.
 29. The method of claim 27, wherein the cell is in a humansubject.
 30. A method of treating a disease or disorder associated withkinase activity in a subject in need thereof, comprising contacting thesubject with a therapeutically effective amount of the compound ofclaim
 1. 31. The method of claim 30, wherein the kinase activity is aJanus kinase activity.
 32. The method of claim 30, wherein the kinaseactivity is a Rho-associated protein kinase activity.
 33. (canceled)